Genetic analyses of ancient tea trees provide insights into the breeding history and dissemination of Chinese Assam tea(Camellia sinensis var.assamica)

Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.

Adaptive mechanisms of Ardisia crenata var.bicolor along an elevational gradient on Gaoligong Mountain,Southwest China

Plants overcome environmental stress by generating metabolic pathways.Thus,it is crucial to understand the physiological mechanisms of plant responses to changing environments.Ardisia crenata var.bicolor has an important ornamental and medicinal value.To reveal the impact of elevational gradient on the habitat soil and plant physiological attributes of this species,we collected root topsoil(0–20 cm)and subsoil(20–40 cm)samples and upper leaves at the initial blooming phase,in a survey of six elevations at 1,257 m,1,538 m,1,744 m,1,970 m,2,135 m,and 2,376 m,with 18 block plots,and 5sampling points at each site.Temperature decreases with an increase in elevation,and soil variables,and enzymatic activities fluctuated in both the topsoil and subsoil,with all of them increasing with elevation and decreasing with soil depth.Redundancy analysis was conducted to explore the correlation between the distribution of A.crenata var.bicolor along the elevational gradient and soil nutrients and enzyme activities,the soil properties were mainly affected by p H at low elevations,and governed by total phosphorus(TP)and available nitrogen(AN)at high elevations.The levels of chlorophyll,carbohydrates,and enzymatic activity except for anthocyanin in this species showed significant variation depending on physiological attributes evaluated at the different collection elevations.The decline in chlorophyll a and b may be associated with the adaptive response to avoid environmental stress,while its higher soluble sugar and protein contents play important roles in escaping adverse climatic conditions,and the increases in activities of antioxidant enzymes except peroxidase(POD)reflect this species’higher capacity for reactive oxygen scavenging(ROS)at high elevations.This study provides supporting evidence that elevation significantly affects the physiological attributes of A.crenata var.bicolor on Gaoligong Mountain,which is helpful for understanding plant adaptation strategies and the plasticity of plant physiological traits along the elevational gradients.

Geographical differences of leaf traits of the endangered plant Litsea coreana Levl.var.sinensis and its relationship with climate

Seventeen morphological and anatomical characteristics of the leaves were selected from five natural populations to explore the variation in leaf traits of Lits ea core ana var.sinensis and the effects of geographical environment on these variations.Nested analysis of variance,multiple comparisons,principal component analysis(PCA),and correlation analysis were conducted to explore the variations within and between populations and their correlation with geographical and climatic factors.Significant differences in the 17 leaf traits were observed within and among populations.On average,the relative contribution of within population variation to total variation was 24.8%,which was lower than among population variation(54.6%).The average differentiation coefficient of the traits was 65.8%,and the average coefficient of variation 11.8%,ranging from6.7%for main vein thickness to 21.4%for petiole length.The PC A results showed that morphological characteristics were divided into two categories,and the level of variation was greater than that of leaf anatomy.Most of the leaf traits were significantly correlated with geography and climate and showed a gradual variation with longitude,latitude,and altitude.In areas with high temperatures,less rainfall,and strong seasonal rainfall,the leaves are larger,longer and thicker.This study shows that variations in leaf traits of L.coreana var.sinensis mainly come from variations among populations.The level of trait differentiation among populations is high and the level of variation within populations low.These findings help further understand leaf morphological characteristics of this species and can provide a valuable reference for the protection and sustainable utilization of this natural resource.

Population dynamics and protection strategies of Litsea coreana Levl.var.sinensis with Guizhou Province in China as an example

Studying the dynamics of plant populations is crucial to the conservation and management of endangered plants.The diameter at breast height(DBH),tree height,crown width,and environmental factors of a total of 15 plots in five populations in Guizhou Province,China were measured in this study.The method of substituting diameter at breast height for age was used to divide the population into 7 age classes.The age structure,static life table,and relationship between the population characteristics and environmental factors of the Litsea coreana Levl.var.sinensis population were studied.Moreover,the time series model was used to predict the population dynamics.Results showed that the age structure of population in Zhengan County(Population ZA)had an inverted Jshape with abundant seedlings and insufficient middle-aged and elderly individuals.The age structures of populations in Daozhen County(Population DZ),Kaiyang County(KY),Meitan County(Population MT),and Xishui County(Population XS)were spindle-shaped with few young individuals and many middle-aged and elderly individuals.The mortality rate of each population fluctuated with the increase in age class,and the populations were extremely unstable.The survival curve was close to the Deevey III type,and the degree of deviation was related to the lack of young individuals.After the age stages II,IV,and VI,the number of aged plants in the different populations initially increased then decreased,and population stability was difficult to maintain.Redundancy analysis showed that the DBH,tree height and crown width of the population decreased with the increase of annual mean temperature,and the population density was restricted by soil nutrient conditions to a certain extent.The results of this work provide scientific basis for the conservation and sustainable utilization of natural populations of L.coreana var.sinensis in Guizhou Province,China.

Effect of Algae on Melon(Cucumis melo subsp.agrestis var.conomon)Growth and Development under Drought-Stress Conditions

Due to climate change,it is necessary to develop plant varieties that are resilient to climate conditions and resistant to abiotic and biotic stresses.The use of microalgae,which are microorganisms that contain carbohydrates,proteins,lipids,and vitamins,against drought tolerance is a new approach.The aim of the current study was to determine the drought-related mechanisms in the conomon melon genotype and develop drought-tolerant melon cultivars.Morphological,physiological,pomological,and molecular analyses were carried out on the algae-treated genotypes.It has been determined that commercial algae application provides the best results in leaf temperature,leaf relative water contents(LRWC),plant height,fruit length,fruit diameter,and yield,while Cag Cag(a special river in the region)stream algae application gives better results in main stem diameter.It was determined that the number of nodes in the control(without algae)plots was higher than in the other two treatments.Yield and LRWC and plant height values of genotype 7 were the best values among other genotypes.The leaf temperature measurement was lower on genotype 9 than on the other genotypes.While the highest fruit length value was measured in genotype 1,genotype 8 was superior in the main stem diameter,fruit diameter,and the number of nodes among the remaining plant materials.In terms of yield,it was determined that the interaction between genotype 2 and the commercial algae resulted in the best outcomes.In addition,the results of the genetic evaluation revealed that the materials used were not genetically distant from each other and more detailed genetic evaluations are needed.The molecular kinship analysis revealed that the genotypes used in the study were divided into three distinct groups,with individuals within each group exhibiting a high degree of relatedness to one another.As a result of the study,it was found that the application of microalgae had significant effects on improving the drought tolerance of Cucumis melo subsp.agrestis var.conomon genotypes.

Quality Standard of Qiang Herb Sophora davidii var.chuansiensis

[Objectives]To establish the quality standard of common clinical folk herb Sophora davidii var.chuansiensis.[Methods]The qualitative identification was performed by microscopic method and thin-layer chromatography(TLC).The contents of moisture,total ash,acid insoluble ash and extract were determined according to the methods of the Chinese Pharmacopoeia(2020).High performance liquid chromatography(HPLC)was used to determine the contents of oxymatrine and oxysophocarpine.[Results]The microscopic characteristics were obvious,including crystal sheath fiber,calcium oxalate square crystal,non glandular hair,stone cells,epidermal cells,stomata,cork cells,vessels with marginal pits and so on.TLC spots were clear,and the resolution was good.The contents of moisture,total ash,acid insoluble ash and extract from 10 batches of samples were 4.70%-8.33%,3.43%-4.19%,0.65%-1.02%and 14.67%-22.04%respectively.The determination results of oxymatrine were between 0.10%-0.33%,with an average value of 0.19%.The determination results of sophocarpine oxide were between 0.30%-0.38%,with an average value of 0.34%.[Conclusions]The established quality standard of S.davidii var.chuansiensis had good specificity and accuracy,and could be used for the quality control of herb S.davidii var.chuansiensis.

Antidepressant-like Effects of Flavonoids from Pteridium aquilinum var.latiusculum

[Objectives]To explore the antidepressant-like effects of PAFL and its potential mechanisms.[Methods]This study used PAFL in mice under various conditions and analyzed the results.The antidepressant-like effects of flavonoids from Pteridium aquilinum(PAFL)and its possible action mechanism were discussed.[Results]PAFL(5 mg/kg,i.p.)exhibited markedly antidepressant-like activities,which could be reversed by treatment with p-chlorophenylalanine(an inhibitor of 5-HT synthesis),haloperidol(a non-selective D 2 receptor antagonist)and N-methyl-D-aspartic acid(an agonist at the glutamate site).Meanwhile,PAFL also effectively increased the hippocampus 5-HT,DA,and Glu levels of mice exposed to TST and FST.The LPS-induced antidepressant behavioral experiment showed that PAFL(1 and 5 mg/kg,i.p.)and fluoxetine(20 mg/kg,i.p.)effectively reduced the immobility time,and increase activity time in OFT.PAFL pretreatment significantly reduced IL-6 and TNF-αserum levels and improve the changes of hippocampal SOD and MDA oxidative stress indicators.Furthermore,PAFL preconditioning could up-regulate BDNF and TrκB contents in hippocampus and down-regulate IκB-αand NF-κB phosphorylation.[Conclusions]The antidepressant-like effects of PAFL might be mediated by the 5-HT,DAergic and Gluergic systems,and the mechanisms of anti-depressant effects of PAFL might be via the alterations of animal behaviors,hippocampus inflammation and neurotrophy,which might be attributed by the BDNF/TrκB/NF-κB signaling pathway.

Effects of Arbuscular Mycorrhizal Fungi on the Physiology and Saponin Synthesis of Paris polyphylla var. yunnanensis at Different Nitrogen Levels

Arbuscular mycorrhizal fungi(AMF)are important members of the plant microbiome and affect the uptake and transfer of mineral elements by forming a symbiotic relationship with plant roots.Nitrogen(N),as an important min­eral element,can directly affect plant growth and development at different N levels.It has been confirmed that inoc­ulation with AMF can improve the efficiency of N utilization by plants.However,there are still fewer reports on the dynamic relationship between arbuscular mycorrhizal and plant secondary metabolites at different nitrogen levels.In this experiment,the physiological indexes and genes related to saponin synthesis were determined by applying differ­ent concentration gradients of nitrogen to the medicinal plant P.polyphylla var.yunnanensis infested by AMF as the test material.It was found that nitrogen addition increased the biomass,chlorophyll content,and nutrient content of above-and below-ground plant parts and increased the content of saponin content of P.polyphylla var.yunnanensis to some extent,but AMF inoculation increased the saponin content of P.polyphylla var.yunnanensis more significantly.AMF inoculation also promoted the expression of genes related to the saponin synthesis pathway,including 3-hy­droxy-3-methylglutaryl coenzyme A synthase(HMGS),squalene epoxidase 1(SE1),and cycloartenol synthase(CAS),which is in according with the accumulation of saponin in plants.It also may increase the saponin content of AMF plants by altering the expression of P450s and UGTs related to saponin synthesis.

光叶苕子(Vicia villosa var.)内生细菌Sz-2的特性及其对香蕉枯萎病的防治效果

【目的】从云南绿肥主推品种光叶紫花苕根内筛选出对香蕉具有促生和拮抗作用的菌株,并对其特性和香蕉枯萎病的防治效果进行研究,以丰富生防促生菌株资源库。【方法】从光叶紫花苕根中筛选出高效拮抗菌株Sz-2,使用平板对峙试验测定菌株抑菌效果;并测定其体外解磷、解钾、固氮及产铁载体能力;通过形态学观察、生理生化测定和16S rDNA序列分析,对该菌株进行了分类鉴定。布置盆栽试验,设4个处理:无抗生菌对照和接种病原菌TR4、Sz-2+TR4、Sz-2,测定香蕉叶片和球茎的发病指数以及促生效果。【结果】光叶紫花苕根内分离得到的菌株Sz-2对香蕉枯萎病菌具有较好的拮抗效果,平板径向抑菌率达74.3%。菌株Sz-2为革兰氏阴性细菌,菌落形态呈浅白色、放射状,接触酶、氧化酶、赖氨酸脱羧酶、β-半乳糖苷酶、精氨酸双水解酶、明胶水解、硝酸还原反应、纤维二糖、柠檬酸反应、葡萄糖发酵反应、酪素水解等均为阳性,吲哚反应、淀粉水解、产H2S反应、鸟氨酸脱羧酶、脲酶反应为阴性。结合形态学和分子生物学鉴定菌株Sz-2为嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)。盆栽结果显示,接种拮抗菌Sz-2后的香蕉球茎和叶片枯萎病发生较轻,球茎纵切面和叶片颜色正常且植株生长健康,对球茎和叶片的防效达69.66%和84.60%;香蕉茎粗、单株鲜重和株高与CK相比分别提高了6.01 mm、83.3 g和11.82 cm。【结论】菌株Sz-2对香蕉枯萎病具有较好的防治作用,加之其解磷、解钾和产铁载体功能,对香蕉植株具有显著的促生效果,可结合绿肥研制具有防控香蕉枯萎病的功能型绿肥,为香蕉促生抗病菌株的进一步田间开发应用提供理论依据。

三峡库区河岸植物野古草(Arundinella anomala var. depauperata Keng)茎通气组织发生对水淹的响应

野古草(Arundinella anomala var. depauperata Keng)在三峡库区长江及其支流江(河)岸有广泛分布,对水淹有很好的耐受能力。有研究表明许多植物在水淹时通气组织发生增强,通气组织的产生改善了植株通气状况,提高了植物对水淹的抵御能力。为了研究水淹是否会影响野古草的通气组织发生以及野古草通气组织发生对水淹的反应,考察了不同水淹深度、不同水淹时间和不同水淹方式处理时野古草茎中通气组织的发生情况。实验中共设置3个水淹深度:不进行水淹(对照)、植株地下部分淹没、植株完全淹没于水下2m深处;5个淹没时间:植株被淹没的时间长度分别为5、10、20、30d和60d;2种水淹方式:连续水淹和间歇水淹。实验结果表明:(1)在无水淹情况下野古草茎中可以产生通气组织,通气组织产生随植株的生长而增强;水淹加快了野古草通气组织发生的进程,促进了野古草通气组织的提前发生。(2)野古草茎中通气组织并不会因为水淹的时间越长而产生越多,植株通气组织的大小达到一定程度后不再因水淹时间的增长而继续增大。(3)淹没深度对通气组织发生有一定影响,总的看来,地下部分淹没野古草植株的通气组织发生要强于完全淹没植株。(4)不同水淹方式对野古草通气组织发生的影响因水淹深度不同而有差异。在完全淹没情况下,连续水淹植株的通气组织比间歇水淹植株的通气组织发达;在地下部分淹没情况下,除水淹初期外,随水淹时间的延长,连续水淹植株通气组织发生与间歇水淹植株没有差异。

Family selection and evaluation of Larix gmelinii var.principis-rupprechtii(Mayr.)Pilger based on stem analysis data at multiple sites

Larix gmelinii var.principis-rupprechtii(Mayr.)Pilger is an important native tree species in North China with advantages of fast growth,straight trunk,and good wood properties.The multi-year and multi-site breeding research of families of the species has not been reported previously.Based on diameter at breast height(DBH),height and volume of 25 families on four experimental sites,we calculated variance components,genetic parameters,juvenile and mature trait correlations and made genotype main effect plus genotype×environment interaction effect(GGE)biplot based on the breeding values estimated using the method of best linear unbiased prediction(BLUP).Compared with height,DBH and volume had higher heritability and larger variation coefficients,making them the more suitable traits for family selection and evaluation.Based on these,GGE biplots containing 20 combinations of site×age were drawn using data at 13 to 17 years when the interactions between family and location were strong.Test sites classifications based on DBH,and volume were inconsistent,with two categories for DBH and one for volume.The Guyuan site was the most suitable with strong discriminating ability,high representativeness and stability among tree ages.Integrating the ranking results of DBH and volume,families 66,76,82 and 111 were high-yielding and stable,families 78 and96 were high-yielding with above average stability,families72 and 79 were high-yielding with below average stability,whereas stability of family 100 was inconsistent between DBH and volume.Early selection based on DBH was convenient and reliable,and can be made at seven years.This study provides support for the selection of Larix gmelinii var.principis-rupprechtii families in Hebei province and an example for the application of stem analysis data from multiple sites in tree breeding.

花椰菜(Brassica oleracea var.botrytis)黑腐病抗性基因同源序列分离及克隆的研究

通过从 NBS 保守序列设计简并引物 PCR 的方法,以花椰菜(Brassica oleracea vat.botrytis)抗、感黑腐病的近等基因系为材料,分离得到 NBS-LRR 型抗性基因同源序列,并获得1个克隆,命名为 RGA330-7.Southern 杂交表明,该片段在近等基因系中存在明显的多态性,且该片段在抗黑腐病基因位点至少存在3个以上类似 RGA330-7的同源拷贝.序列分析结果认为该克隆与 NBS-LRR 型抗性基因的部分 CDSs 有很高的同源性,说明该片段属于 NBS-LRR 型.系统进化分析该序列与甘蓝型油菜的2个抗病同源序列归为一类,很可能这3个不同来源的抗性基因同源序列同属于一种抗性基因家族.因此推测该序列与花椰菜抗黑腐病基因紧密相关,为进一步克隆花椰菜抗黑腐病基因提供了可靠的候选基因,对分子标记辅助抗性育种具有重要意义。

响应面法优化Rhizopus microsporus var.chinensis产生淀粉酶的条件

对Rhizopus microsporus var.chinensis固态发酵产生淀粉酶的条件进行了优化。首先采用单因子实验确定最适固态发酵基质为小麦麸皮,最适碳源和氮源分别为可溶性淀粉和硫酸铵,通过控制培养基初始pH为3.0、初始湿度为70%时,使其生淀粉酶产量提高3.5倍。在此基础上,利用响应面中心组合设计对显著因素进行优化,得出每500mL三角摇瓶中含小麦麸皮13.7g、可溶性淀粉0.063g、硫酸铵0.052g时,生淀粉酶产量达到48.50U/mL,比初始产量提高了8倍。

嗜热子囊菌光孢变种Thermoascus aurantiacus var.levisporusβ-葡萄糖苷酶的分离纯化及特性研究

采用室内培养、测定方法,对嗜热子囊菌光孢变种Thermoascus aurantiacus var.levisporus产生的β-葡萄糖苷酶进行了分离纯化及特性研究。粗酶液经硫酸铵沉淀、DEAE-SepharoseFastFlow阴离子层析、Phenyl-Sepharose疏水层析等步骤获得了凝胶电泳均一的β-葡萄糖苷酶。结果表明,经12%SDS-PAGE测得酶的单亚基分子量约为118kDa,凝胶过滤层析测得酶的分子量约为350kDa。该酶反应的最适温度和最适pH分别为80℃和4.5￣5.0,在pH5.0条件下,该酶在70℃条件下基本稳定,80℃保温30min,剩余酶活为15%。金属离子对β-葡萄糖苷酶活性影响较大,其中Ca2+、Ba2+对酶有激活作用;Ag+、Fe3+、Cu2+对酶有显著的抑制作用。该酶对水杨苷具有很强的底物特异性。

利用Native制备电泳分离来源于Rhizopus microsporus var. chinensis的葡萄糖淀粉酶同工酶

将Native制备电泳应用于Rhizopus microsporus var. chinensis中2个葡萄糖淀粉酶同工酶的分离,考察了电泳缓冲系统、凝胶浓度和凝胶长度对分离效果的影响。结果表明:先使用Ornstein-Davis系统分离同工酶,再使用0.02mol/L,pH6.2的NaAc.CH3COOH缓冲液洗脱,在长度为6cm的7%的Native电泳胶上,2个性质相近仅电泳迁移率略有差别的同工酶能有效分离。分离回收的蛋白质可保持其活力,便于后续研究。

嗜热子囊菌光孢变种Thermoascus aurantiacus var.levisporus内切β-葡聚糖酶的分离纯化及特性

采用培养分离、室内测定等方法,对嗜热子囊菌光孢变种Thermoascus aurantiacus var.levisporus产生的内切β-葡聚糖酶进行了分离纯化及特性研究.粗酶液经硫酸铵沉淀、DEAE-Sepharose Fast Flow阴离子层析、Phenyl-Sepharose疏水层析等步骤获得了凝胶电泳均一的内切β-葡聚糖酶.结果表明,经12%SDS-PAGE测得酶的单亚基分子量约为31.5 kD,凝胶过滤层析测得酶的分子量约为34.5 kD.该酶反应的最适温度为55℃,最适pH为2.5～3.0.该酶在pH3.0条件下60℃较为稳定;80℃保温30 min有20%原酶活性.金属离子对内切β-葡聚糖酶活性影响较大,其中K+、Ca2+、Mn2+对酶有激活作用;Ag+、Cu2+、Al3+对酶有显著抑制作用.该酶对羧甲基纤维素具有很强的底物特异性.

Deciphering codon usage patterns and evolutionary forces in chloroplast genes of Camellia sinensis var. assamica and Camellia sinensis var. sinensis in comparison to Camellia pubicosta

Codon usage bias(CUB) is a unique property of genome which refers to non-random usage of synonymous codons in coding sequences. The present study makes an attempt to find out the pattern of CUB in chloroplast(cp) genes among three tea species, i.e., Camellia sinensis var. assamica(Assam tea), Camellia sinensis var. sinensis(Chinese tea) and Camellia pubicosta(wild tea species) as no work on CUB was reported earlier. To understand the patterns of codon usage among the cp genes of three tea groups, we used bioinformatic tools to investigate the protein coding sequences of cp genes. In our present study, the mean nucleobase T was the highest whereas C was the lowest in all the three tea groups. The overall AT content was more than GC content, i.e., genes were AT rich. The scaled chi-square(SCS) value indicated that the CUB of cp genes was low. The codon CGT(Arg) was over-represented in C. sinensis var. sinensis whereas GGA(Pro) was over-represented in C. pubicosta species. Heatmap study revealed that most of the GC ending codons showed positive correlations between codon usage and GC3 while AT ending codons exhibited negative correlations. From neutrality plot analysis, it was evident that natural selection had played a major role, while mutation pressure exerted a minor effect in the CUB of cp genes in three tea groups. Highly significant(P<0.01) positive correlation was found between SCS and synonymous codon usage order(SCUO) of cp genes which suggested that high expression of cp genes was associated with high degree of CUB.

QTL detection for growth traits in Pinus elliottii var. elliottii and P. caribaea var. hondurensis

Quantitative trait locus （QTL） detection was carried out for growth traits in 122 F1 progenies of Pinus elliottiivar. elliottii （PEE） x tR caribaea var. hondurensis （PCH） hybrid. The PCH male parent linkage map contained 108 mark- ers in 16 linkage groups, while the PEE female parent contained 93 markers in 19 linkage groups. Sequence-related amplified polymorphism （SRAP）, microsatellite （SSR）, expressed sequence tag polymorphism （ESTP） and inter-simple sequence repeat （ISSR） were selected from an existing linkage map. Growth traits investigated were height at age five （HT05） and six years （HT06）, diameter at breast height at age five （DBH05） and six years （DBH06） and annual growth from age five to six years of height （AGHT） and diameter at breast height （AGDBH）. KruskaI-Wallis and interval mapping approaches were used to estimate levels of significance, the number of QTLs, the percentage of the phenotypic varia- tion explained by each of QTLs and their positions on the genetic linkage maps. Twenty six QTLs with significance levels p 〈 0.05 were detected on the parental maps for the six growth traits, which explained more than 15% of the phenotypic variation, suggesting an existence of major-effect genes. Several QTLs had the pleiotropy controlling more than one sin- gle growth trait. Overall, the proportion of phenotypic variation explained by QTLs ranged from 5.9% to 40.6% for HT05 and HT06, from 6.6% to 42.0% for DBH05 and DBH06 and from 5.8% to 22.3% for AGHT and AGDBH. The results from this study provide a basis for marker-aided selection （MAS） in PEE x PCH hybrids.

Metarhizium anisopliae var.anisopliae Pr1A基因的克隆表达及抗血清研究

采用RT-PCR方法从本实验室分离筛选到的金龟子绿僵小孢变种Metarhizium anisopliae vat．anisopliae中,扩增得到PrlA基因全长,此基因全长为1242bp,经Blastn分析此基因序列与M．anopliae的PrlA基因(M73795)同源率为98%。以pET- 22b(+)为基础载体,构建pET-PrlA重组表达载体,在大肠杆菌(Escherichia．coli)BL 21(DE3)中进行表达。经SDS—PAGE分析,获得了约42kDa大小的重组目的蛋白,目的蛋白占表达总蛋白含量的63．2%。将表达的PrlA蛋白切胶回收后制备成抗原,免疫家兔4次后,采血收集抗血清,用ELISA测定效价为1/10000。结果表明,获得的抗体可用于更进一步的研究,将有利于我们进一步了解M．anisopliaeis的侵染机理,弄清楚各Pr蛋白酶的作用方式和对寄主的选择优势,提高生防控制的有效性。

Promoting the Growth of Pinus sylvestris var.mongolica Seedlings and Improving Rhizosphere Fungal Community Structure through Interaction between Trichoderma and Ectomycorrhizal Fungi

In this study,pot experiments were conducted on the seedlings of Pinus sylvestris var.mongolica to study the influence of Trichoderma(Trichoderma harzianum E15)and Ectomycorrhizal fungi(Suillus luteus N94)on the growth of these seedlings.In particular,the effects of these fungi on the fungal community structure in the rhizosphere soil of the seedlings were investigated.Inoculation with Trichoderma harzianum E15 and Suillus luteus N94 significantly(P<0.05)promoted the growth of the Pinus sylvestris seedlings.The non-metric multidimensional scaling(NMDS)results indicated a significant difference(P<0.05)between the fungal community structures in the rhizosphere soil of the annual and biennial seedlings.In the rhizosphere soil of annual seedlings,the main fungi were Ascomycota,Basidiomycota,Zygomycota.Ascomycota,Basidiomycota,Mortierellomycota,and p-unclassified-k-Fungi were the main fungi in the rhizosphere soil of biennial seedlings.The dominant genus in the rhizosphere soil and a key factor promoting the growth of the annual and the biennial seedlings was Trichoderma,Suillus,respectively.Both of them were negatively correlated with the relative abundance of microbial flora in the symbiotic environment.Trichoderma had a significant promoting effect on the conversion of total phosphorus,total nitrogen,ammonium nitrogen,nitrate nitrogen,and the organic matter in the rhizosphere soil of the seedlings,while Suillus significantly promoted the conversion of organic matter and total phosphorus.